Using a sterile 200 µL tip, a small portion of a colony was picked on a Middlebrook 7H10 solid-medium and applied directly on a ground-steel MALDI target plate. Then, one µL of a matrix solution (saturated α-cyano-4- hydroxycinnamic acid in 50% acetonitrile and 2.5% trifluoroacetic acid) (Bruker Daltonics) was used to over-lay the sample. After 5 minutes-drying at room temperature, the plate was loaded into the Microflex LT (Bruker Daltonics) mass spectrometer. Spectra were recorded following the parameters as previously described32 (link). All signals with resolution ≥400 were automatically acquired using AutoXecute acquisition control in flexControl software version 3.0 and the identifications were obtained by MALDI Biotyper software version 3.0 with the Mycobacteria Library v2.0 database (version December, 2015).
Matrix solution
Manufactured by Bruker
The Bruker Matrix Solution is a versatile laboratory equipment designed for a range of analytical applications. It provides a standardized platform for sample preparation, data acquisition, and data analysis. The core function of the Matrix Solution is to facilitate efficient and reliable sample handling and measurement processes in research and industrial laboratories.
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Lab products found in correlation
3 protocols using matrix solution
1
MALDI-TOF MS Identification of Mycobacteria
2
MALDI-TOF Identification of Mycobacteria
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A small part of a colony was picked on a Middlebrook 7H10 solid-medium using a sterile tip and applied directly on a ground-steel MALDI target plate. Then, one µL of a matrix solution (saturated α-cyano-4- hydroxycinnamic acid in 50% acetonitrile and 2.5% trifluoroacetic acid) (Bruker Daltonics) was used to over-lay the sample. After 5 minute-drying, the plate was loaded into the Microflex LT (Bruker Daltonics) mass spectrometer. Spectra were recorded following the parameters as previously described26 . All signals with resolution ≥400 were automatically acquired using AutoXecute acquisition control in flexControl software version 3.0 and the identifications were obtained by MALDI Biotyper software version 3.0 with the Mycobacteria Library v2.0 database (version December, 2015).
3
MALDI-TOF MS Bacterial Protein Profiling
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A loop (approx. 1 μl) of freshly grown bacteria was suspended in ethanol (900 μl, 99.5%), centrifuged at 11,000×g for 2 min, and the pellet was air-dried at room temperature for 3 min. The pellet was incubated in 70% formic acid (20 μl) for 3 min at room temperature to lyse the cells and the released proteins were isolated with acetonitrile (20 μl). The mixture was vortexed and centrifuged at 11,000×g for 2 min. The supernatant was spotted onto a MALDI-TOF MS target plate with Bruker Matrix solution (1 μl). MALDI-TOF MS plates were air-dried at room temperature, and mass spectrometry was performed. Two replicate analyses were performed for each bacterial isolate.
The mass spectra of the bacterial proteins of 2000-20,000 mass-to-charge ratio (m/z) were analyzed using BioNumerics version 7.5 created by Applied Maths NV (http://www. applied-maths.com). Default parameters for strict preprocessing of spectrum data using baseline subtraction, noise elimination, and curve smoothing were selected. Similarity comparison was performed with peak-based Pearson coefficient using default parameters, and phyloproteomic dendrogram was created.
The mass spectra of the bacterial proteins of 2000-20,000 mass-to-charge ratio (m/z) were analyzed using BioNumerics version 7.5 created by Applied Maths NV (http://www. applied-maths.com). Default parameters for strict preprocessing of spectrum data using baseline subtraction, noise elimination, and curve smoothing were selected. Similarity comparison was performed with peak-based Pearson coefficient using default parameters, and phyloproteomic dendrogram was created.
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